【摘要】 目的分子信标探针在芯片表面的固定结合是限制其在芯片技术领域运用的难题。本研究拟寻找一种将分子信标探针固定在芯片表面的新方法,同时对其中重要影响因素进行优化探讨。方法本实验设计一种带单侧延长臂分子信标探针,固定在芯片表面进行杂交、扫描等后续实验。提高杂交效率部分优化包括:分子信标探针浓度;杂交温度、杂交时间;杂交液配方;4种分子对分子信标荧光信号比较。结果较理想区分阴阳性信号强度理想的探针浓度:Cy3-BDH分子信标探针浓度约10μmol/L,FAM-DABCLYE约15μmol/L。讨论单侧延长臂分子信标特异性高、敏感性高、技术难度低,对中、高密度芯片杂交可做到“点对点杂交”。更多还原

【Abstract】 Objective The difficulty of immobilization of molecular beacon probe on the chip has long limited the application of its chip technology.To find a new way of the immobilization of molecular beacon probe on the chip and to optimize the important condition of hybridization.Methods To designe the molecular beacon probe chip with one-side extending arm immobilized on the chip and to carried out hybridization,scan,etc.The optimization included:the molecular beacon probe concentration,temperature,time and compounding of hybridization;fluorescence comparison of four molecular beacons.Results The most ideal probe concentration to distinguish the fluorescence sign between negative and positive is about 10μmol/L(Cy3-BDH) or 15μmol/L(FAM-DABCLYE).Conclusion The specificity and sensitivity of the molecular beacon probe with one-side extending arm are high.The access to technology is easy,and a "point-to-point hybridization" in medium or high density can be obtained.更多还原